Snap frozen tissues were mechanically homogenized using RNase-free pellet pestles (Fisher Scientific, 12-141-368), rinsed in cold PBS, and incubated in cell extraction buffer. Homogenates were incubated on ice for 20 min then centrifuged at 18,000g for 20 min at 4 °C. The supernatants were assayed immediately for levels of apoptotic markers using the Human Bax ELISA Kit (Abcam, ab199080) and Cleaved Caspase-3 DuoSet IC ELISA Kit (R&D Systems, DYC835-2) according to the manufacturers’ protocols. Optical densities were read on a plate reader (Infinite M Nano + , Tecan Group Ltd.) at 450 nm. Raw readouts were analysed using standard curves as references for quantification.
Human bax elisa kit
Manufactured by Abcam
Sourced in United Kingdom
The Human Bax ELISA Kit is a quantitative sandwich enzyme-linked immunosorbent assay (ELISA) designed to measure the levels of the Bax protein in human samples. Bax is a pro-apoptotic member of the Bcl-2 family of proteins and plays a crucial role in the regulation of programmed cell death. The kit utilizes a specific antibody coated on a 96-well plate to capture the Bax protein from the sample, and a detection antibody conjugated with an enzyme for colorimetric quantification. The resulting color intensity is proportional to the amount of Bax present in the sample.
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Lab products found in correlation
2 protocols using human bax elisa kit
1
Apoptosis Markers Quantification
2
Quantifying Cellular Apoptosis via Bax ELISA
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The apoptosis level was measured by testing the Bax level, which was used previously [33] (link). Human Bax ELISA Kit (ab199080) was provided by Abcam (Cambridge, UK). Brie y, cells were lysed in RIPA lyse buffer with protease inhibitor. 50 µL sample was added to the wells, then Antibody Cocktail was added to the wells, followed by the incubation at room temperature for 1 hour. The plate was washed properly and the TMB solution was added to the plate. After 15 min, the stop solution was added to stop the reaction.
OD at 450 nm was measured.
OD at 450 nm was measured.
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